Glycation is a physiologic process that has been implicated in aging. More specifically, it is a non-enzymatic reaction between reducing sugars and the free amino acid groups of proteins, ultimately producing AGE related proteins such as carboxymethyllysine, pentosidine and methyl glyoxal-lysine dimer. In addition to being implicated in a number of diseases, including diabetes, AGE related proteins have associated with skin aging, including progressive changes characterized by decreased tensile strength, increased resistance to enzymatic digestion and increased non-enzymatic cross-links (Pageon et al). Methyl glyoxal and other reactive carbonyl species have also been implicated in disruption of melanocyte (Wondrak et al) and mitochondrial function (Rosca et al) See also, S. Daniel et al., “Collagen glycation and skin aging” accessed on Aug. 8, 2007 at www.mibellebiochemistry.com/pdfs/Collagen_glycation_and_skin_aging_-_CT—2002.pdf.
Mechanistically, methylglyoxal reacts with amino acid residues of proteins, in particular arginyl residues, forming AGE related proteins. G S Gillis, Selwood et al, Takahash et al, Cheung et al., Riley et al, Ahmed et al, Aboro et al, Westwood et al. The glyoxalase system mediates the conversion of the α-ketoaldehyde methylglyoxal to a hydroxyacid. More specifically, the glyoxalase system is composed of two enzymes, glyoxalase I (E.C. 4.4.1.5, lactoylglutathione lyase) (Glo-I) and glyoxalase II (E.C. 3.1.2.6, hydroxyacylglutathione hydrolyase) (Glo-II) and requires a catalytic quantity of reduced glutathione (GSH). In a first non-enzymatic reaction, the Glo-I hemithioacetal is formed from α-ketoaldehyde and reduced GSH. In a second reaction, Glo-I is isomerized to α-D-hydroxyacid thioester which then undergoes enzymatic hydrolysis in the presence of a Glo-II catalyst to α-D-hydroxyacid. Reduced GSH is recycled in the reaction.
Studies report a positive correlation between the amount of AGE in human skin and increasing age. As measured by collagen-linked fluorescence, AGE levels increase at a rate of about 3.7% per year. Skin auto-fluorescence has been correlated with the increased presence of specific AGE. For example, carboxymethyl lysine and pentosidine have been shown to increase five-fold between the ages of 20 and 85.
Surprisingly and unexpectedly, extracts of Reishi mushroom hydrolyzed by an acid protease of Rhizomucor miehei and then rendered substantially devoid of acid-protease activity have been found to upregulate expressions of genes that code for Glo-I. Without wishing to be bound by a theory, applicants believe that by increasing the levels of Glo-I, the levels of α-ketoaldehydes—including, in particular, methylglyoxal—are reduced, thereby decreasing the formation of AGE related protein in mammals.
The Reishi mushroom (Ganoderma lucidum), also known as Red Reishi, Ling Zhi (in China), Yeong Ji (in Korea), has long been used traditional medicine of Asian cultures. See, e.g., S. Aung, “The Clinical Use of Mushrooms from a Traditional Chinese Medical Perspective” International Journal of Medicinal Mushrooms,” Vol. 7, No. 3, pp. 375-376 (2005); P. Poucheret et al. “Biological and Pharmacological Activity of Higher Fungi: 20-Year Retrospective Analysis” Cryptogamie Mycologie, Vol. 27, No. 4, pp. 311-333 (2006).
Reishi has been used an ingredient in oral nutritional supplements. See, e.g., U.S. Patent Application Publication 2003/0008048 teaching a dietary nutritional supplement for helping the body resist the effects of the aging process comprising: (i) at least one of Vitamin c, schisandra berry, raspberry, strawberry, pomegranate, and elderberry; (ii) bilberry or blueberry; (iii) green tea or dark chocolate; (iv) mixed carotenoids, echinacea, or goldenseal; (v) tocopherols, tocotrienols, zinc, or selenium; (vi) Coenzyme Q10; and (vii) reishi mushroom extract.
The use of Reishi in orally-administered medicinal formulations is further described in the patent literature. See, e.g., U.S. Pat. No. 6,541,0431 (treatment of Attention Deficit/Hyperactivity Disorder with a composition containing reishi in combination with Vitamins (A, B5, B6, B12, C and E), reishi, lecithin, choline, 5-hydroxytryptophan and/or tyrosine).
It is known to those of skill in the art that using different solvents can produce different extracts, with different biological activities. In the case of Reishi, an alkali-extracted peptidoglycan from Korean Ganoderma lucidum is reported to be different in chemical composition than a water-based extract of the same plant. The alkali-based extract was reported to contain 6.9% protein and 75.9% carbohydrates (primarily glucose and mannose), with the polysaccharides covalently bound to a polypeptide core. J Y Cheong et al., “Characterization of Alkali-extracted Peptidoglycan from Korean Ganoderma lucidum” Arch. Pharm. Res. Vol. 22, No. 5, pp. 515-519 (1999). The biological activity of the alkali extract is not reported in the Cheong (1999) reference.
An ethanol-extract of the mycelium of Ganoderma lucidum indigenous to the southern part of the Indian subcontinent has been reported to have topical anti-inflammatory effects in animals. See, B. Lakshmi et al., “Antiperoxidative, anti-inflammatory, and antimutagenic activities of ethanol extract of the mycelium of Ganoderma lucidum occurring in South India” Teratog. Carcinog. Mutagen. Vol. 23, Suppl. 1, pp. 85-97 (2003) (intraperitoneal administration of extract reduced inflammation and edema in mice).
U.S. Patent Application Publication No. 2003/0095981 teaches a process for preparing a biologically-active fraction of Ganoderma lucidum having an optical absorbance of from about 200 nm to about 280 nm and use of the active fraction in treating rheumatoid arthritis and osteoarthritis.
The antioxidant and immunomodulatory effects of orally-ingested Ganoderma lucidum have been reported in the scientific literature. See, e.g., J M Lin et al. “Radical scavenger and antihepatotoxic activty of Ganoderma formosanum, Ganoderma lucidum and Ganoderma japonicum” Journal of Ethnopharmacology Vol. 47, No. 1, pp. 33-41 (1995); Z B Lin “Anti-tumor and immunoregulatory activities of Ganoderma lucidum and its possible mechanisms” Acta. Pharmacol. Sin. Vol. 25, No. 11, pp. 1387-95 (November 2004).
Reishi extract has been used in topically-applied skincare products. Origins Plantidote™ Mega-Mushroom Face Serum comprises Reishi in combination with two other mushrooms (Hypsizygus ulmarius (elm oyster) and Cordyceps sinensis) and Reishi. Estée Lauder Re-Nutriv Revitalizing Comfort Créme contains Reishi in combination with wolfberry and ginseng.
Active Concepts (South Plainfield, N.J.) sells a fermented combination of Reishi and Shiitake Mushrooms under the tradename ACB Mushroom Extract SM (INCI name Lactobacillus/Ganoderma Lucidum (Reishi Mushroom) Extract/Lentinus Edodes (Shiitake Mushroom) Extract Ferment Filtrate). According to a Technical Data Sheet (“TDS”), twice-daily application of a quick-break emulsion containing the fermented Reishi/Shiitake complex at a concentration of 5% increased skin hydration and firmness. The same TDS reported increased skin cell turnover from an aqueous solution containing 10% ACB Mushroom Extract SM.
U.S. Patent Application Publication No. 2006/0045894 teaches the use of ACB Mushroom Extract SM in a topically-applied product for reducing sagging or wrinkles in the neck of humans comprising capsaicinoid at stimulates increased localized circulation in the skin. At Paragraph [0031], the mushroom complex is taught to be an optional ingredient, at use concentrations of from 0.05% to 10%.
Arch Personal Care (South Plainfield, N.J.) also sells a combination of Reishi and Shiitake Mushrooms under the tradename NAB® Mushroom Extract (INCI Name Algae Extract & Lentinus Edodes Extract & Ganoderma Lucidum (Reishi Mushroom) Stem Extract). According to the technical information from Arch, skin firmness and hydration were observed to improve after twelve weeks of twice-daily application of a cream containing NAB® Mushroom Extract at a 5% concentration. The Arch TDS likewise reported an increase in skin cell turnover from an aqueous solution containing 10% NAB® Mushroom Extract.
U.S. Pat. No. 7,060,286 teaches topical use of lipids extracted from broken spores of Ganoderma lucidum extracted by a supercritical fluid-carbon dioxide. Claimed uses of the lipid spores are reduction in wrinkles and pigment lightening.
Japanese Patent Application JP4009325 entitled “Beautifying and Whitening Cosmetic” teaches the use of an extract of Ganoderma lucidum with one or more of ascorbic acid, retinol, pyridoxine, pantothenic acid, and tocopherol.
U.S. Patent Application Publication No. 2006/0029686 (assigned to Access Business Group) teaches administration rosehips and at least one of blackberry, blueberry, resveratrol, and extract of Aframomum melegueta to mediate the physiological response caused by interleukin cytokines, specifically inflammation. Primarily directed at oral administration of nutritional supplements containing the above-listed active ingredients, the '686 application teaches compositions additionally containing Reishi.
U.S. Patent Application Publication No. 2005/0158258 (assigned to Mary Kay) teaches topical compositions for treating damaged skin comprising (i) ximenynic acid, a conjugated, unsaturated fatty acid found in sandalwood seeds and (ii) niacin, alpha-lipoic acid, or a mushroom extract. Among the disclosed mushroom extracts is Reishi.
U.S. Patent Application Publication No. 2004/0057917 teaches topical compositions useful in the treatment of photodamaged skin. The disclosed compositions comprise a pharmaceutically-effective amount of an extract of nopal cactus in combination with an anti-inflammatory agent selected from a group including Reishi.
U.S. Patent Application Publication No. 2005/0137239 (assigned to Avon) teaches topical administration of thiazole derivatives to inhibit the formation of advanced glycation endproducts, break advanced glycation endproduct-associated crosslinks, and inhibit glucose oxidase.
U.S. Patent Application Publication No. 2002/0042438 (assigned to L'Oréal) discloses a method for reducing or inhibiting the glycation of skin proteins by topically applying a composition containing ergothioneine and derivatives thereof.
U.S. Patent Application Publication No. 2006/0045896 (assigned to Tracie Martyn Intl) teaches a method for inhibiting glycation of skin proteins and formation of AGE related proteins by topical application of a composition comprising benfotiamine and pyridoxamine.
U.S. Pat. No. 7,005,148 (assigned to L'Oréal) teaches reducing or inhibiting glycation of skin proteins by topical administration of a composition comprising an extract of a Vaccinium-type plant.
U.S. Pat. No. 6,414,038 (assigned to L'Oréal) teaches the use of topical compositions comprising a hydroxystilbene compound to reduce or inhibit the glycation of proteins of the skin, the nails and the hair.
A series of related patents assigned to Active Organics, LP—U.S. Pat. Nos. 5,976,556; 6,569,437; and 6,656,701—describe the uses of one or more acid protease enzymes in combination with an acidic buffering system that enhances epidermal exfoliation and/or epidermal cell renewal, thereby improving the texture or appearance of the skin.
Extracts of Rhizomucor miehei are commercially-available from a number of sources including and Novozymes Inc. and Active Organics LP (Lewisville, Tex.) exhibit enzymatic activity, principally from-acid proteases.
U.S. Patent Application Publication No. 2007/0160563 discloses topical compositions comprising extracts of R. miehei that are substantially devoid of acid-protease activity and their use in treating dermatologic conditions, including reducing the appearance of signs of skin aging.
There has been and remains a need for compositions and methods for reducing or inhibiting the formation of AGE related proteins. The compositions and methods of the present invention—topical application of an effective amount of an extract of reishi mushroom that has been hydrolyzed by an acid protease and thereafter rendered substantially devoid of acid-protease activity—meet this need.